LOCUS       sequence-370800-        6487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Citrine2 for C-terminal tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6487)
  AUTHORS   Storer ISR, Sastre-Velasquez LE, Easter T, Mertens B, Dallemulle A, 
            Bottery M, Tank R, Offterdinger M, Bromley MJ, van Rhijn N, Gsaller 
            F
  TITLE     Shining a light on the impact of antifungals on Aspergillus 
            fumigatus subcellular dynamics through fluorescence imaging.
  JOURNAL   Antimicrob Agents Chemother. 2024 Nov 6;68(11):e0080324. doi: 
            10.1128/aac.00803-24. Epub 2024 Oct 15.
  PUBMED    39404344
REFERENCE   2  (bases 1 to 6487)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Antimicrob 
            Agents Chemother."; date: "2024-11-6"; volume: "68(11):e0080324. 
            doi"; pages: " 10.1128/aac.00803-24. Epub 2024 Oct 15"
COMMENT     Sequence Label: pIN3 (gpdA-cTerm-Citrine2-ptrA)
FEATURES             Location/Qualifiers
     source          1..6487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(120..137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(291..879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(371..390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1050..1841,1842..1910))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1673..1692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1911..2015)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2083..2101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2139..2161)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2261..2280
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2474..2496
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2488..2505
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2489..2505
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2506..2562
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             complement(2875..2886)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(3284..3620,3679..4325)
                     /codon_start=1
                     /gene="ptrA of Aspergillus oryzae"
                     /product="pyrithiamine resistance protein (Kubodera et al.,
                     2000)"
                     /label=ptrA
                     /note="useful as a dominant selectable marker in 
                     Aspergillus"
                     /translation="MSPPAAIYEPTVAATGLKGKVVVSETVPVEGASQTKLLDHFGGKW
                     DEFKFAPIRESQVSRAMTRRYFEDLDKYAESDVVIVGAGSCGLSTAYVLAKARPDLKIA
                     IVEASVSPGGGAWLGGQLFSAMVMRRPAEVFLNELGVPYEEDANPNYVVVKHASLFTST
                     LMSKVLSFPNVKLFNATAVEDLITRPTENGNPQIAGVVVNWTLVTLHHDDHSCMDPNTI
                     NAPVIISTTGHDGPFGAFCAKRLVSMGSVDKLGGMRGLDMSSAEDAIVKNTREVTKGLI
                     IGGMELSEIDGFNRMGPTFGAMVLSGVKAAEEALKVFDERQRECAE"
     primer_bind     4720..4739
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     intron          5054..5169
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     primer_bind     complement(6376..6392)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6376..6392)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6389..6411)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6400..6416
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6424..6430,6431..6448,6449..6454))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(6469..6487,1..3)
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
       61 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
      121 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      181 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
      241 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      301 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      361 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      421 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      481 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      541 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      601 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      661 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      721 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      781 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      841 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
      901 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
      961 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1021 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1081 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1141 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     1201 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1261 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1321 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1381 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1441 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1501 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1561 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1621 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1681 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1741 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1801 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1861 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1921 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1981 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2041 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2101 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     2161 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     2221 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     2281 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     2341 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     2401 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     2461 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt
     2521 acccggggat cctctagagt cgacctgcag gcatgcaagc ttgccaattg attacgggat
     2581 cccattggta acgaaatgta aaagctagga gatcgtccgc cgatgtcagg atgatttcac
     2641 ttgtttcttg tccggctcac cggtcaaagc taaagaggag caaaaggaac ggatagaatc
     2701 gggtgccgct gatctatacg gtatagtgcc cttatcacgt tgactcaacc catgctattt
     2761 aactcaaccc ctccttctga accccaccat cttcttcctt ttcctctcat cccacacaat
     2821 tctctatctc agatttgaat tccaaaagtc ctcggacgaa actgaacaag tcttcctccc
     2881 ttcgataaac ctttggtgat tggaataact gaccatcttc tatagttccc aaaccaaccg
     2941 acaatgtaaa tacactcctc gattagccct ctagagggca tacgatggaa gtcatggaat
     3001 acttttggct ggactctcac aatgatcaag gtatcttagg taacgtcttt ggcgtgggcc
     3061 ggtgttcgtt cccagtcatc gatgcattca catgccctcc ctaagctggg ccctagactc
     3121 taggatccta gtctagaagg acatggcatc gatggactgg gttcgttctg agattatacg
     3181 gctaaaactt gatctggata ataccagcga aaagggtcat gccttctctc gttcttcctg
     3241 ttgatggaat ggctaacaga tgatagacat tgcaacttga aacatgtctc ctccagctgc
     3301 catctacgaa cccactgtgg ccgctaccgg cctcaagggt aaggtcgtgg tttctgagac
     3361 cgtccccgtt gagggagctt ctcagaccaa gctgttggac catttcggtg gcaagtggga
     3421 cgagttcaag ttcgccccta tccgcgaaag ccaggtctct cgtgccatga ccagacgtta
     3481 ctttgaggac ctggacaagt acgctgaaag tgacgttgtc attgttggtg ctggttcctg
     3541 cggtctgagc actgcgtacg tcttggccaa ggctcgtccg gacctgaaga ttgctatcgt
     3601 cgaggccagc gtctctcctg gtcagtagtc catgatggat tgccttgcac tcagctttcc
     3661 ggaactaacg tgcaataggt ggcggtgcct ggttgggtgg ccaactcttt tctgctatgg
     3721 tcatgcgccg tcccgcggaa gtcttcctga acgagctggg tgttccttac gaagaggacg
     3781 caaaccccaa ctacgttgtc gtcaagcacg cctccctgtt tacctcgaca ctcatgtcga
     3841 aggttctctc cttccccaat gtcaagctct tcaatgctac cgctgttgag gacttgatca
     3901 cccgtccgac cgagaacggc aacccccaga ttgctggtgt tgtcgtcaac tggacgctgg
     3961 tcacccttca ccacgatgat cactcctgca tggaccccaa cactatcaac gctcctgtca
     4021 tcatcagtac cactggtcac gatgggccat tcggcgcctt ctgtgcgaag cgcttggtgt
     4081 ccatgggcag cgtcgacaag ctaggtggca tgcgtggtct cgacatgagc tcggccgagg
     4141 atgccatcgt caagaacacc cgcgaggtta ctaagggctt gataatcggc ggtatggagc
     4201 tgtctgaaat tgatggcttt aaccgcatgg gccctacctt cggtgccatg gttctcagtg
     4261 gtgtcaaggc tgccgaggag gcattgaagg tgttcgacga gcgtcagcgc gagtgtgctg
     4321 agtaattact tgtagagctc gtccatgccg agagtgatgc cggcggcggt gacgaactcg
     4381 aggaggacca tgtggtcacg cttctcgttg gggtccttgg agagggcgga ctggtaggag
     4441 aggtagtggt tgtcggggag gaggacgggg ccgtcgccga tgggggtgtt ctgctggtag
     4501 tggtcggcga gctggacgga gccgtcctcg atgttgtgac ggatcttgaa gttgaccttg
     4561 atgccgttct tctgcttgtc ggccatgatg tagacgttgt gggagttgta gttgtactcg
     4621 agcttgtggc cgaggatgtt gccgtcctcc ttgaagtcga tgcccttgag ctcgatacgg
     4681 ttgacgaggg tgtcgccctc gaacttgacc tcggcacggg tcttgtagtt gccgtcgtcc
     4741 ttgaagaaga tggtacgctc ctggacgtag ccctcgggca tggcggactt gaagaagtcg
     4801 tgctgcttca tgtggtcggg gtaacgggcg aagcacatga ggccgtagcc gaaggtggtg
     4861 acgagggtgg gccaggggac ggggagcttg ccggtggtgc agatgaactt gagggtgagc
     4921 ttgccgtagg tggcgtcgcc ctcgccctcg ccggagacgg agaacttgtg gccgttaacg
     4981 tcgccgtcga gctcgacgag gatggggacg acgccggtga agagctcctc gcccttggag
     5041 accatggtga tgtctgctca agcggggtag ctgttagtca agctgcgatg aagtgggaaa
     5101 gctcgaactg aaaggttcaa aggaataagg gatgggaagg atggagtatg gatgtagcaa
     5161 agtacttact taggggaaat aaaggttctt ggatgggaag atgaatatac tgaagatggg
     5221 aaaagaaaga gaaaagaaaa gagcagctgg tggggagagc aggaaaatat ggcaacaaat
     5281 gttggactga cgcaacgacc ttgtcaaccc cgccgacaca ccgggcggac agacggggca
     5341 aagctgccta ccagggactg agggacctca gcaggtcgag tgcagagcac cggatgggtc
     5401 gactgccagc ttgtgttccc ggtctgcgcc gctggccagc tcctgagcgg cctttccggt
     5461 ttcatacacc gggcaaagca ggagaggcac gatatttgga cgccctacag atgccggatg
     5521 ggccaattag ggagcttacg cgccgggtac tcgctctacc tacttcggag aaggtactat
     5581 ctcgtgaatc ttttaccaga tcggaagcaa ttggacttct gtacctaggt taatggcatg
     5641 ctatttcgcc gacggctata cacccctggc ttcacattct ccttcgctta ctgccggtga
     5701 ttcgatgaag ctccatattc tccgatgatg caatagattc ttggtcaacg aggggcacac
     5761 cagcctttcc acttcggggc ggaggggcgg ccggtcccgg attaataatc atccactgca
     5821 cctcagagcc gccagagctg tctggcgcag tggcgcttat tactcagccc ttctctctgc
     5881 gtccgtccgt ctctccgcat gccagaaaga gtcaccggtc actgtacaga gctcacgagt
     5941 tcgtcacatt tttctacaaa tggtggaggc ggcggatttt aggctcaagt catgaccctc
     6001 tgggtcactc cagaatcagc taggtcaacg aataaggatg attctatagg aagatccagg
     6061 caccggtcaa ccatgatctg gacagatttg ggagctcggt ataagctctc caccatctta
     6121 ttctgtatag tttaggctta aagtttatcc aggagatgtt gctgaagtcg atttgagtcc
     6181 acttcctcac tggtagctat acgactttga tggtcgttgt aggggctgta ttaggtctcg
     6241 atcaaacaca aatagaatta aatggtactc gagtccactg aaggtgggct tctccgtctt
     6301 ccgtagccgt gccgaaatcc ttacagcttg tgttgtgtga cttttggtta cgccgtcaag
     6361 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6421 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6481 actcaca
//
